[Development of a DNA biochip for detection of known mtDNA mutations associated with MELAS and MERRF syndromes.].
We developed an oligonucleotide biochip for synchronous multiplex detection of 31 known mitochondrial DNA mutations associated with MELAS (Mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes) and MERRF (Myoclonic epilepsy with ragged red fibers). Allele-specific oligonucleotide probes were covalently immobilized on aldehyde modified glass slides, and then hybridized with Cy5-labled DNA fragments amplified from sample DNAs by a multiplex asymmetric PCR (MAP) method. Five patients with MELAS, 5 patients with MERRF and 20 healthy controls were investigated using the oligonucleotide biochip. The results showed that all the cases with MELAS had an A3243G mutation in the MT-TL1 gene. In the MERRF group, 4 cases were found to be an A8344G mutation and 1 case was a T8356C mutation, and both mutations were in the MT-TK gene. In the healthy controls, none of the 31 related mutations was found. The results of the DNA biochip were consistent with those by DNA sequencing. Clearly, the DNA biochip combined with MAP method would become a valuable tool in multiplex detecting of the point mutations in mtDNA leading to MELAS and/or MERRF syndrome. Moreover, this biochip format could be modified to extend to the screening scope of SNPs for any other human mitochondrial diseases.
Chen
Gang
G
1. Key Lab of Gene Resource Utilization for Severe Hereditary Diseases of Ministry of Education < Key Lab of Genome Research of Anhui Province, Anhui Medical University, Hefei 230032, China; 2. Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China E-mail: gang_7717@hotmail.com.
Li
Wei
W
DU
Wei-Dong
WD
Cao
Hui-Min
HM
Tang
Hua-Yang
HY
Tang
Xian-Fa
XF
Sun
Zhong-Wu
ZW
Zhao
Hui
H
Jin
Qing-Hui
QH
Zhao
Jian-Long
JL
Zhang
Xue-Jun
XJ
D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000235:genetics; D006801:Humans; D017241:MELAS Syndrome / Q000235:genetics; D017243:MERRF Syndrome; D009154:Mutation; D017354:Point Mutation
D004272:DNA, Mitochondrial